Fig. 7
m6 A modification is involved in the upregulation of POU6F2-AS1 in CRC. (A) The m6A modification sites in POU6F2-AS1 were predicted by the SRAMP website. (B, C) The expression of METTL3 and IGF2BP2 between CRC tissues and normal samples in TCGA database. (D, E) Correlation of POU6F2-AS1 and METTL3 or IGF2BP2 expression in CRC was analyzed by GEPIA 2 website. (F) Representative images of POU6F2-AS1, METTL3 and FASN analyzed by FISH or IHC in POU6F2-AS1 high expression and POU6F2-AS1 low-expression tumours. (G) The correlation between POU6F2-AS1 and METTL3 was analyzed in CRC tissues using the FISH score and IHC score. (H) After transfecting shRNAs of METTL3 into SW480 cells, the mRNA expression of METTL3 and POU6F2-AS1 was verified by qRT–PCR. (I) MeRIP-qPCR analysis of the m6A levels in POU6F2-AS1 in SW480 cells transfected with the METTL3 shRNAs. (J) The stability of POU6F2-AS1 treatment by actinomycin D at the indicated times following METTL3 knockdown or overexpression in CRC cells was examined by qRT–PCR. (K) After transfecting siRNAs of IGF2BP2 into SW480 cells, the mRNA expression of IGF2BP2 and POU6F2-AS1 was verified by qRT–PCR. (L) The stability of POU6F2-AS1 treatment by actinomycin D at the indicated times following IGF2BP2 knockdown in SW480 cells was examined by qRT–PCR. (M) RIP-qPCR showing the enrichment of IGF2BP2 on POU6F2-AS1 in SW480 cells with METTL3 knockdown. (N) Schematic diagram of the mechanism by which the m6A modification-mediated POU6F2-AS1 lncRNA facilitates lipogenesis and growth in CRC